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α-Glucosidase inhibitory activity and protease characteristics produced by Bacillus amyloliquefaciens (Bacillus amyloliquefaciens로부터 생산된 protease 특성 및 α-glucosidase 저해활성)

  • Lee, Rea-Hyun;Yang, Su-Jin;Hwang, Tae-Young;Chung, Shin-Kyo;Hong, Joo-Heon
    • Food Science and Preservation
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    • v.22 no.5
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    • pp.727-734
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    • 2015
  • In this study, three GRAS (generally recognized as safety) strain was isolated from Doenjang and Cheonggukjang and identified as a protease-producing microorganism, following the appearance of a clear zone around its colony when cultured on a medium containing skim milk. Based on an analysis of the nucleotide sequence of 16S ribosomal RNA, the strains wereas identified as Bacillus amyloliquefaciens and wereas therefore named Bacillus amyloliquefaciens CDD5, Bacillus amyloliquefaciens CPD4, and Bacillus amyloliquefaciens CGD3. Here, we analyzed the protease and ${\alpha}$-glucosidase inhibitory activities of the three B. amyloliquefaciens strains. Among the isolated strains, B. amyloliquefaciens CGD3 exhibited the highest protease activity (9.21 U/mL, 24 hr). The protease activities of B. amyloliquefaciens CDD5 and B. amyloliquefaciens CPD4 reached 1.14 U/mL and 8.02 U/mL, respectively, at 48 hr. The proteases from the three B. amyloliquefaciens strains showed the highest activities within a pH range of 8.0-9.0 at $50^{\circ}C$, and casein was found to be the preferred substrate on evaluating enzyme activity in the substrate specificity assay. The B. amyloliquefaciens strains exhibited maximal growth when the nutrient broth medium had an initial pH within the range of 5.0-10.0, 6-9% sodium chloride (NaCl), and 5% glucose. B. amyloliquefaciens CDD5 exhibited a low ${\alpha}$-glucosidase inhibition rate (5.32%), whereas B. amyloliquefaciens CPD4 and B. amyloliquefaciens CGD3 exhibited relatively higher inhibition rates of 96.89% and 97.55%, respectively.

Production and fermentation characteristics of seafood kimchi started with Leuconostoc mesenteriodes SK-1 isolated from octopus baechu kimchi (문어 배추김치에서 분리한 Leuconostoc mesenteroides SK-1을 이용한 수산물김치의 제조 및 발효 특성)

  • Jang, Mi-Soon;Jung, Ko-Eun;Yun, Jae-Ung;Nam, Ki-Ho
    • Food Science and Preservation
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    • v.23 no.7
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    • pp.1050-1057
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    • 2016
  • This study was carried out to investigate the effectiveness of Leuconostoc mesenteroides isolated from octopus baechu kimchi as a potential starter for seafood kimchi. L. mesenteroides is lactic acid bacterium currently used as a starter for kimchi production. We selected the most effective L. mesenteroides strain from the 7 strains isolated from octopus baechu kimchi and, based on biochemical properties and 16S rRNA sequencing, identified the selected strain as L. mesenteroides SK-1. The SK-1 strain exhibited acid-tolerance, good survival capacity, and excellent dextran productivity. We investigated the effects the SK-1 of starter on seafood kimchi fermentation. Octopus baechu kimchi was fermented with L. mesenteroides SK-1 at $4^{\circ}C$ for 35 d. The decrease in pH and increase in acidity in octopus baechu kimchi fermented with the SK-1 starter occurred more quickly than that in the control kimchi indicating that. Octopus baechu kimchi with SK-1 starter has a relatively slow rate of increase in lactic acid production. As a result, octopus baechu kimchi prepared with L. mesenteroides SK-1 can be maintained at a suitable ripening degree over an extended period of time compared to that of the control kimchi, Moreover, the octopus baechu kimchi started with L. mesenteroides SK-1 has excellent sensory properties, including a refreshing taste, and a weak sour odor.

Batch Scale Storage of Sprouting Foods by Irradiation Combined with Natural Low Temperature - I. Storage of Potatoes - (방사선(放射線) 조사(照射)와 자연저온(自然低溫)에 의한 발아식품(發芽食品)의 Batch Scale 저장(貯藏)에 관(關)한 연구(硏究) - 제(第) 1 보(報) : 감자의 저장(貯藏) -)

  • Cho, Han-Ok;Byun, Myung-Woo;Kwon, Joong-Ho;Yang, Ho-Sook;Lee, Chul-Ho
    • Korean Journal of Food Science and Technology
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    • v.14 no.4
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    • pp.355-363
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    • 1982
  • In order to develop the commercial storage method of potatoes by irradiation combined with natural low temperature, storage room($450{\times}650{\times}250cm$; year round temperature change, $2-17^{\circ}C;\;70-85%\;R.H.$) on a batch scale followed by irradiation with optimum dose level. Irish cobbler and Shimabara were 100% sprouted after 3 months storage in control, whereas in 15 Krad irradiated group, sprouting was completely inhibited at Irish cobbler for 9 months storage, and at Shimabara for 12 months. The extent of loss due to rot attack after 9 months storage was 6% in control, 6-8% in 10-15 Krad irradiated group at Irish cobbler and weight loss was 16.5% in control, 5.1-5.6% in irradiated group, whereas rotting rate of Shimabara after 12 months storage was 100% in control, 15% in irradiated group and the weight loss of its was 12.6% in control, $7.3{\sim}7.4%$ in irradiated group. The moisture content in whole storage period of two varieties were $72{\sim}82%$ without remarkable changes. The total sugar and ascorbic acid contents were slightly decreased according to the dose increase and elapse of storage period, whereas reducing sugar content was increased. Irish cobbler was 90% marketable after 9 months storage and 85% in Shimabara after 12 months storage.

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Apoptotic Effects and Cell Cycle Arrest Effects of Extracts from Cnidium monnieri (L.) Cusson through Regulating Akt/mTOR/GSK-3β Signaling Pathways in HCT116 Colon Cancer Cells (HCT116 대장암세포에서 AKT/mTOR/GSK-3β 신호경로 조절을 통한 벌 사상자 추출물(CME)의 apoptosis 및 cell cycle arrest 효과)

  • Lim, Eun Gyeong;Kim, Guen Tae;Kim, Bo Min;Kim, Eun Ji;Ha, Sung Ho;Kim, Sang-Yong;Kim, Young Min
    • Journal of Life Science
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    • v.26 no.6
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    • pp.663-672
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    • 2016
  • The Cnidium monnieri (L.) Cusson is an annual plant distributed in China and Korea. The fruit of C. monnieri is used as a medicinal herb that is effective for the treatment of carbuncle and pain in female genitalia. However, the anti-cancer effects of CME have not yet been reported. In this study, we assessed the apoptotic effects and cell cycle arrest effects of ethanol extracts from C. monnieri on HCT116 colon cancer cells. The results of an MTT assay and LDH assay demonstrated a decrease in cell viability and the cytotoxic effects of CME. In addition, the number of apoptotic body and the apoptotic rate were increased in a dose-dependent manner through Hoechst 33342 staining and Annexin V-PI double staining. In addition, cell cycle arrest occurred at the G1 phase by CME. Protein kinase B (Akt) plays an important role in cancer cell survival, growth, and division. Akt down-regulates apoptosis-mediated proteins, such as mammalian target of rapamycin (mTOR), p53, and Glycogen Synthase kinase-3β (GSK-3β). CME could regulate the expression levels of p-Akt, p-mTOR, p-GSK-3β, Bcl-2 family members, caspase-3, and PARP. Furthermore, treatment with CME, LY294002 (PI3K/Akt inhibitor), BIO (GSK-3β inhibitor), and Rapamycin (mTOR inhibitor) showed that apoptotic effects occurred through the regulation of the AKT/mTOR/GSK-3β signaling pathway. Our results demonstrated CME could induce apoptosis and cell cycle arrest in HCT116 colon cancer cells.

Enhanced Production of Carboxymethylcellulase by a Newly Isolated Marine Microorganism Bacillus atrophaeus LBH-18 Using Rice Bran, a Byproduct from the Rice Processing Industry (미강을 이용한 해양미생물 Bacillus atrophaeus LBH-18 유래의 carboxymethylcellulase 생산의 최적화)

  • Kim, Yi-Joon;Cao, Wa;Lee, Yu-Jeong;Lee, Sang-Un;Jeong, Jeong-Han;Lee, Jin-Woo
    • Journal of Life Science
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    • v.22 no.10
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    • pp.1295-1306
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    • 2012
  • A microorganism producing carboxymethylcellulase (CMCase) was isolated from seawater and identified as Bacillus atrophaeus. This species was designated as B. atrophaeus LBH-18 based on its evolutionary distance and the phylogenetic tree resulting from 16S rDNA sequencing and the neighbor-joining method. The optimal conditions for rice bran (68.1 g/l), peptone (9.1 g/l), and initial pH (7.0) of the medium for cell growth was determined by Design Expert Software based on the response surface method; conditions for production of CMCase were 55.2 g/l, 6.6 g/l, and 7.1, respectively. The optimal temperature for cell growth and the production of CMCase by B. atrophaeus LBH-18 was $30^{\circ}C$. The optimal conditions of agitation speed and aeration rate for cell growth in a 7-l bioreactor were 324 rpm and 0.9 vvm, respectively, whereas those for production of CMCase were 343 rpm and 0.6 vvm, respectively. The optimal inner pressure for cell growth and production of CMCase in a 100-l bioreactor was 0.06 MPa. Maximal production of CMCase under optimal conditions in a 100-l bioreactor was 127.5 U/ml, which was 1.32 times higher than that without an inner pressure. In this study, rice bran was developed as a carbon source for industrial scale production of CMCase by B. atrophaeus LBH-18. Reduced time for the production of CMCase from 7 to 10 days to 3 days by using a bacterial strain with submerged fermentation also resulted in increased productivity of CMCase and a decrease in its production cost.

Effects of Reed (Phragmites communis) on Growth Performance, Carcass Characteristics, and Meat Quality in Hanwoo Steers (갈대 급여가 한우 거세우의 발육, 도체 및 육질특성에 미치는 영향)

  • Lee, Sang Min;Chang, Sun Sik;Chung, Ki Yong;Kim, Hyeong Cheol;Choi, Sun Ho;Jeong, Ha Yeon;Yang, Boh Suk;Lee, Sung Sill;Cho, Young Moo
    • Journal of Life Science
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    • v.22 no.10
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    • pp.1392-1398
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    • 2012
  • This study investigated the effects of diet of different forages on growth performance and carcass characteristics of Hanwoo steers. Twenty-one Hanwoo steers were randomly allocated to three groups (fed hay, reed, and reed with rice straw) of seven steers each. Initial and final body weights of control, T1, and T2 groups were 125.5, 128.3, 128.3 kg and 697.4, 614.6, 706.7 kg, respectively. Average daily gain tended to increase in controls (0.70 kg/d) and the T2 group (0.71 kg/d) but not as much in the T1 group (0.60 kg/d); however, there was no significant difference. DMI was not significantly different among the treatment groups, but T1 was relatively lower than the other groups. For the yield traits, carcass weight was not significantly different between controls and the T2 group but was greater in the T2 group compared to the T1 group (p<0.05). Back fat thickness and rib eye area were higher in controls and T2 compared to T1; yield grade (A:B:C, %) was greater in T1 (43:57:0) compared to the other groups (control 0:71:29; T2 29:42:29). For the quality traits, fat color and texture were not significantly different among groups. However, meat color and maturity were significantly greater in T1 compared to T2 (p<0.05). Marbling score and appearance rate of over 1st meat quality grade were greater in the control and T2 groups compared to the T1 group. Based on the results, growth performance, feed utilization, and carcass traits appeared to improve when roughage containing rice straw plus reed was offered. Therefore, reed is worth considering as a roughage source for fattening Hanwoo steers.

Long-term Climate Change Research Facility for Trees: CO2-Enriched Open Top Chamber System (수목의 장기 기후변화 연구시설: CO2 폭로용 상부 개방형 온실)

  • Lee, Jae-Cheon;Kim, Du-Hyun;Kim, Gil-Nam;Kim, Pan-Gi;Han, Sim-Hee
    • Korean Journal of Agricultural and Forest Meteorology
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    • v.14 no.1
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    • pp.19-27
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    • 2012
  • The open-top chamber (OTC) system is designed for long term studies on the climate change impact on the major tree species and their community in Korea. In Korea Forest Research Institute (KFRI), the modified OTC system has been operating since September 2009. The OTC facility consists of six decagon chambers (10 meters in diameter by 7 meters high) with controlled gas concentration. In each chamber, a series of vertical vent pipes are installed to disperse carbon dioxide or normal air into the center of the chamber. The OTC is equipped with remote controlled computer system in order to maintain a stable and elevated concentration of carbon dioxide in the chamber throughout the experimental period. The experiment consisted of 4 treatments: two elevated $CO_2$ levels ($1.4{\times}$ and $1.8{\times}$ ambient $CO_2$) and two controls (inside and outdoors of the OTC). Average operational rate was the lowest (94.2%) in June 2010 but increased to 98% in July 2010 and was 100% during January to December 2011. In 2010~2011, $CO_2$ concentrations inside the OTCs reached the target programmed values, and have been maintained stable in 2011. In 2011, $CO_2$ concentrations of 106%, 100% and 94% of target values has been recorded in control OTC, $1.4{\times}$ $CO_2$-enriched OTC and $1.8{\times}$ $CO_2$-enriched OTC, respectively. With all OTC chambers, the difference between outside and inside temperatures was the highest ($1.2{\sim}2.0^{\circ}C$) at 10 am to 2 pm. Temperature difference between six OTC chambers was not detected. The relative humidity inside and outside the chambers was the same, with minor variations (0~1%). The system required the highest amount of $CO_2$ for operation in June, and consumed 11.33 and 17.04 ton in June 2010 and 2011, respectively.

Histological studies on in vitro Propagation of Pulsatilla koreana Nakai (할미꽃 기내증식(器內增殖)에 관(關)한 조직학적(組織學的) 연구(硏究))

  • Lee, Man-Sang;Oh, Ki-Hong
    • Korean Journal of Medicinal Crop Science
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    • v.1 no.2
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    • pp.137-157
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    • 1993
  • This study was carried out to investigate the optimal condition for multiple propagation through leaf tissue culture and to apply anther culture techniques to Pulsatilla koreana Nakai breeding. Leaf and anther of Pulsatilla koreana Nakai were cultured on MS, MT, LS and $B_5$ media supplemented with several growth regulators and nitrogen sources under various conditions. For callus induction and differentiation from the Pulsatilla koreana leaf segments were more effective in the combination of zeatin and auxin than auxin alone. The color of the callus was green when treated with IBA alone. Shoot differentiation was more effective when treated with zeatin than auxin alone, especially the best hormoal combination for shoot differentiation was zeatin 1.0mg/l +NAA 0.1mg/l, while 2,4-D inhibited shoot differentiation. The appeared rate of S pollen was 35% in vivo, while that of S pollen by low temperature$(4^{\circ}C)$ pretreatment for 4 days was increased by 53% and the optimum culture time for callus induction from anther was uni-nucleate stage. $B_5$ basal medium supplemented with NAA 0.5mg/l and zeatin 1 mg/l was the most effective on callus formation and the best results of plant regeneration were obtained from combination of NAA 0.5mg/l and zeatin 0.5mg/l in anther culture. $NH_{4}NO_3$ as more effectives as the nitrogen source than $KNO_3$ and the combination with zeatin 2.0mg /L was the best effective. The best combination for plant regeneration in callus induced from anther was $NH_{4}NO_3$ 1650mg/l + $KNO_3$ 3800mg/l + zeatin 2.0mg/l. Ploidy level of anther-derived plants appeared 28% haploid, 47% diploid and the others were triploid, tetraploid and mixploid. In compare with E.S.T, M.D.H and P.X banding patterns were distinguished among callus, haploid and diploid plants in electrophoresis.

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Trial for Drug Susceptibility Testing of Mycobacterium tuberculosis with Live and Dead Cell Differentiation (세포 염색 방법을 이용한 결핵균 감수성 검사법)

  • Ryu, Sung-Weon;Kim, Hyun-Ho;Bang, Mun-Nam;Park, Young-Kil;Park, Sue-Nie;Shim, Young-Soo;Kang, Seongman;Bai, Gill-Han
    • Tuberculosis and Respiratory Diseases
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    • v.56 no.3
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    • pp.261-268
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    • 2004
  • Background : The resurgence of tuberculosis and outbreaks of multidrug resistant (MDR) tuberculosis have increased the emphasis for the development of new susceptibility testing of the Mycobacterium tuberculosis for the effective treatment and control of the disease. Conventional drug susceptibility testings, such as those using egg-based or agar-based media have some limits, such as the time required and difficulties in determining critical inhibitory concentrations, but these are still being used in many diagnostic laboratories because of no better lternatives, considering cost and accuracy. To overcome these limits, a rapid and simple method for new susceptibility testing, using live and dead assays, was applied for a bacterial cell viability assay to distinguish dead from live bacterial cells based on two-color fluorescence. Materials and Methods Strains : Forty strains were used in this study, 20 susceptible to all antituberculosis drugs and the other 20 resistant to the four first line antituberculosis drugs isoniazid, rifampicin, streptomycin and ethambutol. Antibiotics : The four antibiotics were dissolved in 7H9 broth to make the following solutions: $0.1{\mu}g\;isoniazid(INH)/m{\ell}$, $0.4{\mu}g\;rifampicin(RMP)/m{\ell}$, $4.0{\mu}g\;streptomycin(SM)/m{\ell}$ and $4.0{\mu}g\;ethambutol(EMB)/m{\ell}$. Results : Live and dead Mycobacterium tuberculosis cells fluoresced green and red with the acridin (Syto 9) and propidium treatments, respectively. These results are very well accorded with conventional drug susceptibility testing by proportional method on Lowensen-Jensen media (L-J) containing 4 drugs (INH, RMP, EMB and SM), showing a 93.7 % accordance rate in susceptible strains and 95% in resistant strains. Conclusion : The results of the drug susceptibility testing using the live and dead bacterial cell assay showed high accordance rates compared with the conventional proportion method on L-J. This finding suggests that the live and dead bacterial cell assay can be used as an alternative to conventional drug susceptibility testing for M. tuberculosis strains.

Nutrient Dynamics in Decomposing Leaf Litter and Litter Production at the Long-Term Ecological Research Site in Mt. Gyebangsan (계방산 장기생태조사지의 낙엽 생산량 및 낙엽 분해에 따른 양분 동태)

  • Lee, Im-Kyun;Lim, Jong-Hwan;Kim, Choon-Sig;Kim, Young-Kul
    • Journal of Ecology and Environment
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    • v.29 no.6
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    • pp.585-591
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    • 2006
  • We measured the litterfall quantity and investigated the nutrient dynamics in decomposing litter for three years at the LTER sites installed in a deciduous broadleaf natural forest in Mt. Gyebangsan, South Korea. Litterfall production was significantly different among the sampling dates, whereas it was not significantly different among the years. The total annual mean litterfall production for three years was 6,593 kg $ha^{-1}$ $yr^{-1}$ and leaf litter accounted for 82.6% of the litterfall. The leaf litter quantity was highest in Quercus mongolia, followed by leaf of other species, Betula schmidtii, Kaplopanax pictus, Acer pseudo-sieboldianum, etc., which are dominant tree species in the site. The mass loss from the decomposition of leaf litter was fastest in Cortinus controversa (100%), followed by A. preudo-sieboldianum, K. pictus, and B. schmidtii. 100% of litter for C. controversa, 96.1% for A. pseudo-sieboldianum, 92.8% for K. pictus decomposed, while 66.2% of litter for Q. mongolia decayed for 1,003 days. The lower rate of the mass loss in the litter of Q. mongolia may be attributed to the difference in substrate quality, such as lower nutrient concentrations compared with those of other tree species. The concentrations of N, P, and Ca for five litter types increased over time, while the concentrations of K and Mg decreased over time. Compared with the nutrients in the litter of Q. mongolia, the nutrients (N, P, K, Ca, Mg) in the litter of other species, C. controversa, A. pseudo-sieboldianum, and K. pictus, were released more rapidly. The results showed that the mass loss and the nutrient dynamics in the litter are variable depending on the tree species even in the same site conditions.